Comparative Study of the Transfection Efficiency and Toxicity of 293T Cells Transfected by Three Reagents
CHEN Feng, JIN Xinghan, WANG Fei*
This paper aims to compare the transfection efficiencies of two luciferase reporter genes into human embryonic kidney cells 293T by different transfection reagents. By using Lipofectamine® 2000, X-tremeGENE HP DNA, FuGENE® HD transfection reagents that with different ratios of plasmids DNA and transfection reagents, the plasmids containing GFP (green fluorescent protein) and RFP (red fluorescent protein) were transfected into 293T cells, respectively. The transfection efficiency was analyzed by inverted fluorescence microscope and flow cytometry, and the cell survival rate was detected by trypan blue stain. Regardless of the target gene and ratio of plasmid DNA to transfection reagents, the transfection efficiency of X-tremeGENE HP DNA was significantly higher than Lipofectamine® 2000 (all P<0.05). Under the same ratio of plasmid DNA to transfection reagent, the efficiency of FuGENE® HD mediated GFP was significantly higher than Lipofectamine® 2000 (all P<0.05), but there was no significant difference of RFP transfection between the two reagents (all P>0.05). When the ratio of plasmid DNA to transfection reagent was 1:2, the efficiency of X-tremeGENE HP DNA mediated GFP and RFP transfection was significantly higher than FuGENE® HD reagent (all P<0.05), When the ratio of plasmid DNA to transfection reagent was changed to 1:4, the transfection efficiency between the two reagents was not significantly different (all P>0.05). With the increase of the amount of transfection reagents, the efficiencies of Lipofectamine® 2000 and FuGENE® HD transfection reagent were increased significantly, whereas the transfection efficiency of X-tremeGENE HP DNA transfection reagent was decreased significantly. Compared with GFP gene transfection, the transfection efficiency was significantly decreased with the increase of target gene RFP fragment (all P<0.05). When the ratio of plasmid DNA to transfection reagent was the same, there was no significant difference in the survival rate after transfection among the three reagents (all P>0.05), while when the amount of transfection reagent increased, the survival rate was significantly decreased (all P<0.05). The target gene was inversely proportional to the transfection efficiency. Considering the amount of transfection reagent, transfection efficiency and cell activity, this study believed that the X-tremeGENE HP DNA transfection reagent had the best effect.
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